Determination of tetracycline residues in chicken meat by liquid chromatography-tandem mass spectrometry

Analysis of residual levels of tetracyclines (TCs) in chicken meat was performed using a validated liquid chromatography coupled with a tandem mass spectrometry (LC-MS/MS) technique. Overall, the recoveries for TCs ranged from 56.9% to 101.2%, with standard deviations of 4.5–13.2%. Detection limits ranged from 7.9 to 14.6?µg?kg^?1. In four of 60 samples, doxycycline (DXC) was determined in a range from 19.9 to 35.6?µg?kg^?1; and in one sample tetracycline was detected at 17.2?µg?kg^?1. Chlortetracycline (CTC) and oxytetracycline (OTC) were not detected in any of the tested samples. This study indicates that chicken meat sold in Bursa, Turkey, contained some residues of TCs. Therefore, stricter regulations for the use of antibiotics in the poultry industry and the monitoring of drug residues in chicken meat prior to marketing are needed. Finally, this method has been applied successfully for the confirmation of TCs in chicken meat.     

Pilot survey of hen eggs consumed in the metropolitan area of Rio de Janeiro,Brazil, for polyether ionophores,macrolides and lincosamides residues

A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method, which has recently been developed and validated, was used for the identification and quantification of polyether ionophore, macrolide and lincosamide residues in commercial eggs sold in the metropolitan area of Rio de Janeiro, Brazil. The method was applied to 100 samples and the results showed a high incidence of polyether ionophore residues (25%). Salinomycin was detected in 21% of samples, but only two non-compliant results (5.3 and 53 µg?kg^?1) were found if maximum limits (tolerances) established by European Union were adopted in Brazil and if a method decision limit (CCα) of 3.4?µg?kg^?1 was considered. In 8% of analyzed samples, more than one studied coccidiostat was found. The lincosamide, lincomycin, and the macrolide, tylosin, were detected at trace levels in 4 and 1% of the samples, respectively. Lasalocid, clarithromycin and erythromycin were not found.     

Incidence of tetracycline residues in chicken meat and liver retailed to consumers

The incidence of tetracyclines’ (TCs) residue (oxytetracycline, tetracycline, chlorotetracycline and doxycycline) in fresh chicken samples (meat and liver) collected during one year was recorded. TC residues were analysed using an HPLC-DAD method. The limit of detection for meat was 4.4, 5, 10 and 7?ng?g^?1 for OTC, TTC, CTC and DOC, respectively, compared with 5.5, 6, 12 and 9?ng?g^?1 stated for liver. The recovery of the method ranged from 91% to 70% depending upon both matrix type and tetracycline kind. The results revealed that 66 (44%) samples contained TC residues including 21 (42%) breast, 19 (38%) thigh and 26 (52%) liver samples. The corresponding contaminated ranges were 124–5812, 107–6010 and 103–8148?µg?kg^?1. A total of 12 (8%), 13 (7.33%) and 20 (13.33%) samples of breast, thigh and liver, respectively, had TC residues above the Codex maximum residue limit (MRL). Liver samples had a higher incidence and level than those found in breast or thigh samples.     

Implementation of the Bacillus cereus microbiological plate used for the screening of tetracyclines in raw milk samples with STAR protocol – the problem with false-negative results solved

In antibiotic residue analyses the first step of screening is just as important as the following steps. Screening methods need to be quick and inexpensive, but above all sensitive enough to detect the antibiotic residue at or below the maximum residue limit (MRL). In the case of a positive result, the next step is conducted and further methods are used for confirmation. MRLs stated in European Union Regulation 37/2010 for tetracyclines in raw milk are: 100 µg kg^–1 for tetracycline, 100 µg kg^–1 for oxytetracycline, 100 µg kg^–1 for chlortetracycline and no limit for doxycycline because it is prohibited for use in animals from which milk is produced for human consumption. The current five-plate microbiological screening method for the detection of antibiotic residues in raw milk was found to be simple and inexpensive, but not specific, sensitive and reliable enough to detect tetracycline at MRL in routine raw milk screening procedures. Spiking samples with tetracycline at the MRL level and applying them on Bacillus cereus ATCC 11778 microbiological plates often gave false-negative results, indicating that tetracyclines may have to be inactivated or masked. Tetracyclines seem to bind to a certain component in milk. Consequently, when applying samples to the B. cereus microbiological plate the antibiotic cannot inhibit the growth of B. cereus which disables the formation of inhibition zones on the test plate. After adding the appropriate amount of citric acid into the milk samples, we solved the problem of false-negative results. During the validation 79 samples of milk were spiked with tetracyclines at different concentrations: 100 µg kg^–1 for tetracycline, 100 µg kg^–1 for oxytetracycline, 80 µg kg^–1 for chlortetracycline and 30 µg kg^–1 for doxycycline. Concentrations used in the validation matched the requirements for MRLs (they were either at or below the MRLs) stated in European Union Regulation 37/2010. The sensitivity of the validation was 100%.     

Determination of acrylamide in Thai-conventional snacks from Nong Mon market,Chonburi using GC-MS technique

Acrylamide in Thai-conventional snacks was analysed by GC/MS with a linear response ranged of 5–50?µg and r ^2?>?0.99. The limit of detection (s/n?=?(3) and limit of quantification (s/n?=?10) were 4 and 15?µg?kg^?1, respectively, and RSD?<?2%. Acrylamide in 19 food samples ranged from <15?µg?kg^?1 to 1.26?mg?kg^?1 with highest concentrations in Kanom Jak. Moderate levels (150–500?µg?kg^?1) were detected mostly in deep-fried products, especially sweet potato and taro crisps, Kanom Kai Hong, banana fritters, durian chips and spring rolls. Thai-conventional snacks possessed low concentrations (<150?µg?kg^?1) including Khao Larm, Pa Tong Koo, sweet banana crisps and deep-fried Chinese wonton. Acrylamide was lowest (<15?µg?kg^?1) in fish strips, rice crackers, Hoi Jor and fried fish balls. Dietary habits by 400 tourists indicate a daily intake of acrylamide <150?ng, well below a toxic dose.     

1,3‐Dimethoxybenzene,a newly identified component of port wine

1,3‐Dimethoxybenzene was identified by GC–O, GC–MS and Kovats indices (polar Supelcowax, 1709; non‐polar Rtx‐5MS, 1158) as a new volatile component of port wine. Sensory evaluation described this compound as having a sweet medicinal odour with hazelnut, resinous and woody notes. Respective threshold limits in model wine and port wine were 21 and 47 µg l^?1. Quantitative analysis by GC–MS, using a selected characteristic ion (m/z 138), indicated that young port wines from the 1998 vintage contained up to 3 µg l^?1 whereas ports from the 1999 vintage contained up to 20 µg l^?1. © 2002 Society of Chemical Industry     

A multi-class multi-residue method for the analysis of polyether ionophores,tetracyclines and sulfonamides in multi-matrices of animal and aquaculture fish tissues by ultra-high performance liquid chromatography tandem mass spectrometry

ABSTRACT

An ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) based multiclass multi-residue method for the simultaneous analysis of 5 polyether ionophores, 4 tetracycline and 10 sulfonamides in animal and aquaculture fish tissues was developed and validated. Sample extraction and clean-up were based on a modified QuEChERS method. The method was validated using an in-house validation based on performance characteristics modified from Commission Decision 2002/657/EC. Both matrix effect and uncertainties associated with sample preparation and instrumental analysis were minimised by the use of matrix-matched calibrations. Recoveries of analytes were generally satisfactory and typically fell between 80% and 113%. The repeatability and intermediate reproducibility measured as relative standard deviations were in most cases less than 15% (n = 63). The decision limit (CCα) and detection capability (CCβ) ranged from 110.7 to 125.8 and 121.5 to 151.7 µg kg^?1 for tetracyclines, 113.4 to 118.3 and 116.8 to 126.5 µg kg^?1 for sulfonamides and 50.8 to 52.4 and 51.5 to 55.6 µg kg^?1 for polyether ionophores, respectively. The method displayed its fitness for purpose through satisfactory results obtained in international proficiency testing schemes. The method was applied to animal and aquaculture fish tissues obtained from different sources in South Africa. Polyether ionophores were predominantly detected in samples in the range 4.26–290.10 µg/kg. Oxytetracycline was found in one porcine liver sample; however, none of the targeted analytes were present above the detection limit in the aquaculture samples.     

Survey of aflatoxin M1 in cow's milk for human consumption in Kerman Province of Iran

The aim of this study was to assess levels of aflatoxin M₁ (AFM1) in milk samples from Kerman, Iran. AFM1 was detected in 72 samples, ranging in concentration from <0.01 to 0.41?µg?l^?1. The samples were analyzed using immunoaffinity column for clean-up and HPLC for determining AFM1. Milk samples were collected from six dairy farms. AFM1 was found in ～50% of the milk samples. The average level of AFM1 was below the tolerance limit (0.05?µg?l^?1), but 50% of the samples had greater levels than the maximum tolerance limit accepted by EU and the Iranian national standard. The method detection limit and limit of quantification were 0.01 and 0.03?µg?l^?1, respectively, and recovery of the method was 87%. The results showed that AFM1 contamination is a serious problem for public health. To achieve a low level of AFM1 in milk, cattle feed must be monitored regularly for aflatoxin contamination and protected from fungal contamination as much as possible.     

Oxytetracycline residues in retail chicken eggs in Ibadan,Nigeria

Oxytetracycline residues in chicken eggs from five markets within Ibadan metropolis were determined by high-performance liquid chromatography following solid-phase extraction. Recovery of oxytetracycline was 80.5%–87.8%, whereas 75.2% of the samples contained detectable oxytetracycline with an overall mean residue concentration of 479.0?µg?kg^?1. Residue concentrations from Iwo Road, Apata, Challenge, Ojoo and Bodija markets were 421, 460, 468, 568 and 476?µg?kg^?1, respectively. Eggs from Ojoo market had the highest levels, whereas 68.8% of the samples contained residues above the Codex Alimentarius Commission maximum residue limit. The results are of public health interest as they indicate that a greater proportion of eggs being consumed in Ibadan could contain residues above the maximum residue limit, for which unregulated access and indiscriminate use of antibiotics by poultry farmers could be responsible. National surveillance of eggs and other animal products for antibiotic residue and appropriate regulation of antibiotic is recommended to ensure food safety for consumers.     

Development and validation of multi-residue analysis for tetracycline antibiotics in feed by high performance liquid chromatography coupled to mass spectrometry

ABSTRACT

A new multi-residue method for the analysis of the tetracycline antibiotics oxytetracycline, tetracycline, chlortetracycline, and doxycycline was developed and validated for animal feed. After extraction with 0.1M Na₂EDTA-McIlvaine buffer (pH 4), the samples were centrifuged, purified by SPE (Strata-X-CW cartridges) and analysed by HPLC-MS. Validation of the method was performed according to the guidelines indicated in European Commission Decision 2002/657/EC. The procedure was validated by spiking feed samples at three different levels (300, 1000, and 5000 µg/kg). Average recoveries for tetracyclines were in the range 79.70–98.8%, with RSD for repeatability and reproducibility in the ranges of 5.0–9.5% and 6.5–11.0%, respectively. The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in animal feed. The validated method was successfully applied to 65 suspect feed samples collected from different regions of Poland in 2015–16 and obtained from farms and feed manufactures. Of these 65 purportedly non-medicated feeds, eight (12.3%) were positive for the presence of doxycycline or chlortetracycline.     

Ochratoxin A in wines,musts and grape juices from Spain

A survey on the occurrence of ochratoxin A (OTA) in 240 grape‐based beverages was carried out. Red and white wines from four different Spanish Designations of Origin (n = 160), musts (n = 20), grape juices (n = 10), ordinary wines (n = 20), special wines (Malaga, muscatel, sherry, vermouth, etc) (n = 20) and sparkling wines (n = 10) were assayed for OTA content using immunoaffinity column clean‐up and high‐performance liquid chromatography with fluorimetric detection (detection limit 0.05 µg l^?1). Forty‐three (17.9%) of the samples tested contained detectable levels of OTA. The overall mean OTA concentration in red and white wines of Designations of Origin was 0.30 and 0.18 µg l^?1 respectively (ranges 0.05–3.19 and 0.05–1.13 µg l^?1 respectively). The percentage of wine samples with detectable amounts of OTA was higher for red (18.3%) than for white (10%) wines. OTA was also found in two of 10 red ordinary wines (0.68 and 4.24 µg l^?1), whereas none of 10 white ordinary wines contained OTA. The mean OTA amount detected in sparkling wines was 0.44 µg l^?1 (range 0.14–0.71 µg l^?1). Two of 20 must samples contained OTA at low levels (0.08 and 0.18 µg l^?1), while none of 10 grape juice samples contained OTA. Highest amounts of OTA were found in special wines (45%), with a maximum of 15.25 µg l^?1 in a muscatel sample. Copyright © 2004 Society of Chemical Industry     

Development of a novel solid-phase extraction,LC-MS/MS method for the analysis of ethyl carbamate in alcoholic beverages: application to South African wine and spirits

Ethyl carbamate (EC) is a known genotoxic carcinogen that is frequently present in alcoholic beverages and is therefore a public health concern. As a consequence, maximum concentration levels for EC in these commodities are legislated in several countries. Quantitative analytical methods are therefore essential to monitor EC levels in beverages. Most published analytical methods for the determination of EC in alcoholic beverages utilise elaborate sample pre-treatment procedures to obtain injectable samples, or yield low sensitivity, for example where direct injection is used. In addition, these procedures often require large volumes of toxic solvents and are not generally applicable to diverse alcoholic beverages. This paper describes a novel procedure for the determination of EC in wines, fortified wines and spirits. The procedure is based on reversed-phase solid-phase extraction (SPE) sample clean-up combined with normal-phase liquid chromatography-atmospheric pressure chemical ionisation tandem mass spectrometric (NP-LC-APCI-MS/MS) analysis. This method provides a rapid, robust and simple analytical procedure suitable for the analysis of a diverse range of alcoholic beverages. The accuracy of the method (expressed as average recovery from diverse matrices) is 94.5%, with limits of detection (LODs) ranging between 0.25 and 0.63?µg?l^?1 for different matrices. Benefits such as simplified sample preparation, low detection limits, low solvent consumption and good selectivity render the methodology ideally suited to study the occurrence of EC in diverse commodities. The method was applied to study the occurrence of EC in South African wines, fortified wines and spirits. South African wines, aged 1–9 years, contained 1.8–31?µg?l^?1 EC (RSD?=?69%, n?=?106), fortified wines aged 2–34 years contained 2.8–79?µg?l^?1 EC (RSD?=?89%, n?=?21), and brandies aged 3–20 years contained 4.4–95?µg?l^?1 EC (RSD?=?105%, n?=?26). Factors affecting the formation of EC in these commodities were investigated and storage temperature, alcohol content and pH were found to affect the rate of EC formation. Of these variables storage temperature has by far the greatest effect.     

Natural occurrence of type-B trichothecene mycotoxins in Korean cereal-based products

Type-B trichothecenes (deoxynivalenol (DON), nivalenol (NIV), fusarenone-X (FUS-X), 15-acetyldeoxynivalenol (15ADON), and 3-acetyldeoxynivalenol (3ADON)) were determined in 338 cereal-based products. Detection limit, quantification limit and mean recovery for five toxins were in the ranges 0.7–2.6?µg?kg^?1, 2.1–7.8?µg?kg^?1 and 73–110%, respectively. The range of occurrence and average level in samples were, respectively, 21–88% and 5.2–121.8?µg?kg^?1 for NIV, 10–96% and 1.7–109.5?µg?kg^?1 for DON, 2–39% and 0.4–3.6?µg?kg^?1 for FUS-X, 0–80% and 0–17.3?µg?kg^?1 for 15ADON, and 0–29% and 0–1.5?µg?kg^?1 for 3ADON. Regarding co-occurrence, 64% of samples had more than two type-B trichothecenes. The estimated daily intakes of NIV, DON, FUS-X, 15ADON, and 3ADON were 0.077, 0.048, 0.004, 0.006 and 0.002?µg?kg^?1?bw?day^?1, respectively. These results suggest that current exposure levels do not indicate the possibility of adverse effects, but consideration of the combined exposure of type-B trichothecenes may be required due to the high frequency of co-occurrence.     

Multi-residue method for the confirmation of four avermectin residues in food products of animal origin by ultra-performance liquid chromatography–tandem mass spectrometry

A confirmatory method was developed for the rapid determination of abamectin, ivermectin, doramectin and eprinomectin residues in various food products of animal origin, such as pork muscle, pork liver, fish and milk. Samples were homogenized, extracted and de-proteinized by acetonitrile, cleaned via two-step cleaning procedure using Bond Elut C₁₈ SPE columns and then alumina-N cartridges. All the four avermectin residues in different animal-food products were simultaneously separated and determined by ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) within 3.5?min. Data acquisition under positive ESI–MS/MS was performed by applying multiple reaction monitoring (MRM) for both identification and quantification, and mass spectrometric conditions were optimized to increase selectivity and sensitivity. The matrix-matched calibration curves for different matrices, such as pork muscle, pork liver, fish and milk, were constructed and the interference effect of different sample matrices on the ionization was effectively eliminated. The UPLC–MS/MS method was validated with satisfactory linearity, recovery, precision and stability. Matrix-matched calibration curves of abamectin, ivermectin, doramectin and eprinomectin in four different matrices were linear (r^{2 ?}≥?0.990, goodness-of-fit coefficients ≤12.8%) in the range 2.5–200?µg?kg^?1. The limits of detection and quantification for the four avermectins were in the range 0.05–0.68 and 0.17–2.27?µg?kg^?1, respectively. Recoveries were 62.4–104.5% with good intra- and inter-day precision. The method was rapid, sensitive and reliable, and can be applied to the quantitative analysis of avermectin residues in different animal-food products.     

Primary aromatic amine migration from polyamide kitchen utensils: method development and product testing

A rapid and sensitive LC-MS/MS method for the identification, quantification and confirmation of six primary aromatic amines (PAAs) was developed and validated to ISO 17025:2005. From a literature survey, 57 frequently used PAA compounds were identified and subsequently reduced to six – aniline, 4,4′-MDA, 3,3′-DMB, 2,4-TDA, 2,6-TDA and o-T – based on results from migration studies on a range of utensils. Low LOQs of between 0.075 and 0.496?µg?l^?1 were determined for the six analytes, thereby quantifying well below the legal limit of 10?µg?kg^?1 total PAAs. Furthermore, low measurement uncertainties were calculated for the analytical method, in the range of 3.15–3.20%. Mean recoveries were between 98% and 102% and spanned over ±12% at 95% CI. Following the analysis of 84 black polyamide kitchen utensils, the migration of PAAs detected was significant and is therefore of concern. The six analytes identified, quantified and confirmed in this survey could be utilised as possible markers for the identification of PAA migration, thereby improving the time and cost-efficiency of food control laboratories.     

Determinative and confirmatory method for residues of tetracyclines in honey by LC-MS/MS

A limited number of substances are authorised for the treatment of bees. Maximum residue limits (MRLs) are set for tetracyclines in several matrices, but not for honey. Nevertheless, tetracycline antibiotics may be used in order to prevent bacterial diseases and the loss of honey bee populations. In this study, a sensitive multi-residue LC-MS/MS method was developed and optimised for the quantitative and qualitative determination of tetracycline residues in honey. Homogenisation of samples under acidic conditions was performed and solid-phase extraction was carried out. The eluate was evaporated under nitrogen and dissolved in an aqueous methanol solution prior to filtration. A mobile phase composed of acetic acid–water and acetic acid–acetonitrile was used. Separation of tetracycline, oxytetracycline, chlortetracycline and doxytetracycline was achieved by using gradient elution on a C18 chromatography column. The analytical method was validated according to Commission Decision 2002/657/EC by the analysis of spiked samples around the recommended concentration of 20 μg kg^?1 by EURL Guidance Paper, December 2007. A matrix effect was observed, so quantification was based on an external matrix calibration curve. Calculated decision limits (CCα) were lower than 10 μg kg^–1 for all tetracyclines. Good linearity, repeatability and within-laboratory reproducibility were achieved.     

Assessment of average exposure to organochlorine pesticides in southern Togo from water,maize (Zea mays) and cowpea (Vigna unguiculata)

Drinking water, cowpea and maize grains were sampled in some potentially exposed agro-ecological areas in Togo and analysed for their contamination by some common organochlorine pesticides. A total of 19 organochlorine pesticides were investigated in ten subsamples of maize, ten subsamples of cowpea and nine subsamples of drinking water. Analytical methods included solvent extraction of the pesticide residues and their subsequent quantification using gas chromatography-mass spectrometry (GC/MS). Estimated daily intakes (EDIs) of pesticides were also determined. Pesticides residues in drinking water (0.04–0.40 µg l^?1) were higher than the maximum residue limit (MRL) (0.03 µg l^?1) set by the World Health Organization (WHO). Dieldrin, endrin, heptachlor epoxide and endosulfan levels (13.16–98.79 µg kg^?1) in cowpea grains exceeded MRLs applied in France (10–50 µg kg^?1). Contaminants’ levels in maize grains (0.53–65.70 µg kg^?1) were below the MRLs (20–100 µg kg^?1) set by the Food and Agriculture Organization (FAO) and the WHO. EDIs of the tested pesticides ranged from 0.02% to 162.07% of the acceptable daily intakes (ADIs). Population exposure levels of dieldrin and heptachlor epoxide were higher than the FAO/WHO standards. A comprehensive national monitoring programme on organochlorine pesticides should be undertaken to include such other relevant sources like meat, fish, eggs and milk.     

Survey of aflatoxins in maize tortillas from Mexico City

In Mexico, maize tortillas are consumed on a daily basis, leading to possible aflatoxin exposure. In a survey of 396 2-kg samples, taken over four sampling days in 2006 and 2007 from tortilla shops and supermarkets in Mexico City, aflatoxin levels were quantified by HPLC. In Mexico, the regulatory limit is 12?µg?kg^?1 total aflatoxins for maize tortillas. In this survey, 17% of tortillas contained aflatoxins at levels of 3–385?µg?kg^?1 or values below the limit of quantification (<LOQ) and, of these, 13% were >12?µg?kg^?1 and 87% were below the regulatory limit. Average aflatoxin concentrations in 56 contaminated samples were: AFB₁ (12.1?µg?kg^?1); AFB₂ (2.7?µg?kg^?1); AFG₁ (64.1?µg?kg^?1) and AFG₂ (3.7?µg?kg^?1), and total AF (20.3?µg?kg^?1).     

Determination of sterigmatocystin in cheese by high-performance liquid chromatography-tandem mass spectrometry

Different cheese samples produced in Latvia (eight) and Belgium (13) were analysed for their sterigmatocystin (STC) content. Only two (9.5%) of the samples were positive for STC with concentration levels of 1.23 and 0.52 µg kg^?1, respectively. Five (24%) samples contained STC above the limit of detection (0.03 µg kg^?1) but below the limit of quantification (0.1 µg kg^?1), A sensitive liquid chromatography-electrospray positive ionisation-tandem mass spectrometry (LC-MS/MS) method, which was previously developed for the analysis of STC in grains, was modified and applied to the analysis of STC in cheese. This method involved sample extraction with acetonitrile–water (90 : 10, v/v), defatting with n-hexane, solid-phase extraction, separation on a reversed-phase C₁₈ column, and STC detection by LC-MS/MS.     

Herbicide contamination of live armclad rockfish,clam and pen shell by moss-control agents used in aquariums of seafood restaurants in Korea

Determination of simazine and diuron by high-performance liquid chromatography-ultraviolet detection (HPLC-UV) in moss-control agents, seawater and fish in aquariums was investigated and validated. The detection limits are 0.2 (simazine) and 0.4?µg l^–1 (diuron) in blank seawater, and 0.20 (simazine) and 0.30 µg kg^?1 (diuron) in blank fish homogenate, while the recoveries ranged from 93.9% to 102.4% with a relative standard deviation?≤?9.8% for simazine and diuron. The method was successfully used in the study of simazine and diuron contamination in live fish stored in seafood restaurant aquaria in Korea. It was found that 0.4–2.3% of simazine and <0.10–3.8% of diuron were included the in moss-control agents tested. Of the 66 sampled aquarium seawaters, simazine was found to be present in four samples (3.8–42?µg?l^?1) while diuron was detected in two samples (1.3–1.6?µg?l^?1). For fish homogenates used in a bioconcentration study, simazine content ranges from 0.17 to 1.8?mg?kg^?1.